Abstract
15N is a nonradioactive heavy isotope of nitrogen, widely used for biochemical and physiological research in plants. For instance, 15N-KNO3 was used as the nitrogen source in plants in order to investigate nitrate uptake activity and transport from roots to shoots (Lin et al., 2008). Here, we describe a detailed pipeline used for labeling living rice (Oryza sativa) plants with 15N-KNO3 and determination of net nitrate uptake and transport activity, and this protocol was proved to be valid in Arabidopsis and rice (Lin et al., 2008; Hu et al., 2015).
Keywords: Nitrate uptake, Nitrate transport, Rice, Isotope
Materials and Reagents
Equipment
Procedure
Representative data
Figure 1. A 96-well plate that was cut off the bottom well-suited for the growth of rice seedlings. After being put into the 96-well plate, rice roots could grow downward into solution underneath while rice shoots could grow upward tidily along each well. Figure 2. 10-day-old rice seedlings grown on a 96-well plate. The container under the 96-well plate is full of modified Kimura B solution, usually the container is wrapped with light-tight material (e.g., tinfoil) to protect rice roots from light. Table1. 15N content of shoots and roots Table2. Calculation of 15N-nitrate uptake activity and root-to-shoot transport activity
Notes
Recipes
Acknowledgments
The hydroponic culture has been illuminated in Lin et al. (2008) and Hu et al. (2015), the 15N-nitrate uptake activity and root-to-shoot transport activity assay was cited from Hu et al. (2015). This work was supported by grants from the Ministry of Science and Technology of China (2014AA10A602-5, 2015CB755702), and the Chinese Academy of Sciences (XDA08010400).
References
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